Transfection of HL-60 cells by Venus lentiviral vector.
10.7534/j.issn.1009-2137.2013.03.008
- Author:
Zheng LI
1
;
Shao-Yan HU
;
Jian-Nong CEN
;
Zi-Xing CHEN
Author Information
1. Department of Hematology, The Affiliated Children Hospital of Soochow University, Jiangsu Province, China.
- Publication Type:Journal Article
- MeSH:
Genetic Vectors;
HL-60 Cells;
Humans;
Lentivirus;
genetics;
Transfection
- From:
Journal of Experimental Hematology
2013;21(3):576-580
- CountryChina
- Language:Chinese
-
Abstract:
In order to study the potential of Venus, lentiviral vector, applied to acute myeloid leukemia, the recombinant vector Venus-C3aR was transfected into 293T packing cells by DNA-calcium phosphate coprecipitation. All virus stocks were collected and transfected into HL-60, the GFP expression in HL-60 cells was measured by flow cytometry. The expression level of C3aR1 in transfected HL-60 cells was identified by RT-PCR and flow cytometry. The lentiviral toxicity on HL-60 was measured by using CCK-8 method and the ability of cell differentiation was observed. The results indicated that the transfection efficacy of lentiviral vector on HL-60 cells was more than 95%, which meets the needs for further study. C3aR1 expression on HL-60 cells increased after being transfected with recombinant lentiviral vector. Before and after transfection, the proliferation and differentiation of cells were not changed much. It is concluded that the lentiviral vector showed a high efficacy to transfect AML cells and can be integrated in genome of HL-60 cells to realize the stable expression of interest gene. Meanwhile, lentiviral vector can not affect HL-60 cell ability to proliferate and differentiate.
